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文献和实验Western Blotting with Monoclonal Ant
the blocking buffer from the blot, add the antibody solution, and incubate with agitation for 30 minutes at 37˚C, one hour at room temperature, or overnight at 4˚C. Enzyme conjugated secondary antibody Note: The inclusion of sodium azide is to be avoided
Antibody Purification: Ion-Exchange Chromatography
Ion exchange chromatography techniques are the focus of this chapter and they showcase the power of this method for the purification of proteins and monoclonal antibodies. The technique is powerful and can separate biomolecules
Western Blotting with Monoclonal Antibodies
lysis buffer (1% SDS, 1.0mM sodium ortho-vanadate, 10mM Tris pH 7.4). Microwave for 10?5 seconds. Centrifuge the homogenate (16,000 x g, 15 C) for 5 minutes to pellet insoluble material, then discard pellet. Dilute an aliquot
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